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Proliferation of seeded urothelial and smooth muscle cells on different collagen scaffolds and commercially available small intestinal submucosa
ESPU Meeting 2007
S57
# S10-4 (PP)
PROLIFERATION OF SEEDED UROTHELIAL AND SMOOTH MUSCLE CELLS ON DIFFERENT COLLAGEN SCAFFOLDS AND COMMERCIALLY AVAILABLE SMALL INTESTINAL SUBMUCOSA Luc ROELOFS, Jody NUININGA, Herman VAN MOERKERK*, Willeke DAAMEN*, Egbert OOSTERWIJK, Toin VAN KUPPEVELT* and Wout FEITZ Radboud University Nijmegen Medical Centre, Urology, Nijmegen, NETHERLANDS - * Radboud University Nijmegen Medical Centre, Biochemistry, Nijmegen, NETHERLANDS
PURPOSE Different scaffolds have been used for bladder wall regeneration. In search for better materials we have developed collagen scaffolds and compared the invitro viability and proliferation of seeded urothelial and smooth muscle cells on 3 different collagen scaffolds and the commercially available small intestinal submucosa (SIS).
collagen and SIS scaffolds, and cultured under standard conditions. After 3, 7, 14 and 21 days cell viability and proliferation was analysed with the WST-1 assay. Univariate ANOVA test was used for statistical analysis. Seeded scaffolds were paraffin embedded and analyzed by H&E, cytokeratins AE1/AE3, E-cadherin, asmooth muscle actin and desmin staining.
layered smooth muscle cells on top of all scaffolds. Minimal penetration of cells into the scaffolds occurred exclusively in the collagen scaffolds. SIS scaffolds still contained porcine cell-nuclei, as previously reported.1 The collagen scaffolds were completely clear of native cellular remnants.
RESULTS CONCLUSIONS
MATERIAL AND METHODS The collagen scaffolds were created of bovine collagen type I, and chemically cross-linked: 1) porous scaffold; 2) duallayer scaffold, combining a film-layer with a porous layer; 3) porous scaffold with one side closed. Urothelial cells (SCaBER cell line) and smooth muscle cells (PM151T cell line) were separately seeded on the
Progressive growth of seeded cells was observed on all scaffolds. Collagen scaffolds showed better proliferation and viability of cells in comparison to SIS (p < 0.001). No statistically significant differences existed between the different collagen scaffolds. Histology and immunohistochemistry demonstrated the formation of continuous multi-layered urothelial and single-
Based on in-vitro cell proliferation, collagen scaffolds are better devices for cell growth than the SIS scaffold. The collagen scaffolds are better defined and not contaminated with cellular remnants. 1 Feil et al. Investigations of urothelial cells seeded on commercially available small intestinal submucosa. European Urology 2006; June 16
# S10-5 (O)
THE FATE OF THE MUCOSA IN BLADDERS AUGMENTED WITH DEMUCOSALIZED SIGMOID COLON; LONG TERM HISTOLOGICAL RESULTS Pedro-jose LOPEZ E., Ricardo ZUBIETA, Nelly LETELIER, Jose Manuel ESCALA, Gabriela RETAMAL, Gustavo PUIATTI and Samuel BENVENISTES* Hospital Exequiel Gonzalez Cortes, Paed Urology, Santiago, CHILE - * Hospital Exequiel Gonzalez Cortes, Pathology Depart, Santiago, CHILE
PURPOSE Bladder augmentation (BA) using demucosalized sigmoid colon (DSC) is associated with a decrease in mucus production. The aim of this study is to determinate the histological changes at various times after augmentation.
MATERIAL AND METHODS 24 specimens were taken from 20 patients who had undergone BA using DSC over 7 years (1998-2005). Biopsy was done in 3 different points of the augmented patch (9-12-3 o’clock). In children with a closed bladder neck a trans-vesical trochar was used. Histopathology (HþE) results were compared at different post augment
periods. The biopsies were compared with 2 cases of BA where sigmoid colon without demucosalisation was used (SCWD)(>10 years).
with atrophic glands. In the group >36 months the patch was mostly covered with intestinal mucosa with atrophic glands surrounded by fibrotic tissue and no signs of urothelium. In the 2 cases of BA with SCWD the intestinal glands were normal.
RESULTS The mean age at biopsy was 12 years (range 4e15y) with a mean post BA of 39 months (range 12e84m). On endoscopy there were 2 groups; one (<3 years) where the intestine was covered by urothelium with some small areas of intestinal mucosa (‘‘islets’’) (n¼9), and the other >3 years where the patch was covered completely with intestinal mucosa (n ¼ 15). The histology of the early biopsies (<36 months) showed urothelium with some small ‘‘islets’’ of sigmoid mucosa, but
CONCLUSIONS This study has demonstrated regrowth of sigmoid colon mucosa, which may begin from intestinal mucosa remnants (‘‘islets’’), noted on specimens taken <3 years after BA. However the glandular structure is atrophic in those patches, and was normal in those cases augmented with non-demucosalized sigmoid. This may explain the reduced mucus production in those patients augmented with DSC.
S57
# S10-4 (PP)
PROLIFERATION OF SEEDED UROTHELIAL AND SMOOTH MUSCLE CELLS ON DIFFERENT COLLAGEN SCAFFOLDS AND COMMERCIALLY AVAILABLE SMALL INTESTINAL SUBMUCOSA Luc ROELOFS, Jody NUININGA, Herman VAN MOERKERK*, Willeke DAAMEN*, Egbert OOSTERWIJK, Toin VAN KUPPEVELT* and Wout FEITZ Radboud University Nijmegen Medical Centre, Urology, Nijmegen, NETHERLANDS - * Radboud University Nijmegen Medical Centre, Biochemistry, Nijmegen, NETHERLANDS
PURPOSE Different scaffolds have been used for bladder wall regeneration. In search for better materials we have developed collagen scaffolds and compared the invitro viability and proliferation of seeded urothelial and smooth muscle cells on 3 different collagen scaffolds and the commercially available small intestinal submucosa (SIS).
collagen and SIS scaffolds, and cultured under standard conditions. After 3, 7, 14 and 21 days cell viability and proliferation was analysed with the WST-1 assay. Univariate ANOVA test was used for statistical analysis. Seeded scaffolds were paraffin embedded and analyzed by H&E, cytokeratins AE1/AE3, E-cadherin, asmooth muscle actin and desmin staining.
layered smooth muscle cells on top of all scaffolds. Minimal penetration of cells into the scaffolds occurred exclusively in the collagen scaffolds. SIS scaffolds still contained porcine cell-nuclei, as previously reported.1 The collagen scaffolds were completely clear of native cellular remnants.
RESULTS CONCLUSIONS
MATERIAL AND METHODS The collagen scaffolds were created of bovine collagen type I, and chemically cross-linked: 1) porous scaffold; 2) duallayer scaffold, combining a film-layer with a porous layer; 3) porous scaffold with one side closed. Urothelial cells (SCaBER cell line) and smooth muscle cells (PM151T cell line) were separately seeded on the
Progressive growth of seeded cells was observed on all scaffolds. Collagen scaffolds showed better proliferation and viability of cells in comparison to SIS (p < 0.001). No statistically significant differences existed between the different collagen scaffolds. Histology and immunohistochemistry demonstrated the formation of continuous multi-layered urothelial and single-
Based on in-vitro cell proliferation, collagen scaffolds are better devices for cell growth than the SIS scaffold. The collagen scaffolds are better defined and not contaminated with cellular remnants. 1 Feil et al. Investigations of urothelial cells seeded on commercially available small intestinal submucosa. European Urology 2006; June 16
# S10-5 (O)
THE FATE OF THE MUCOSA IN BLADDERS AUGMENTED WITH DEMUCOSALIZED SIGMOID COLON; LONG TERM HISTOLOGICAL RESULTS Pedro-jose LOPEZ E., Ricardo ZUBIETA, Nelly LETELIER, Jose Manuel ESCALA, Gabriela RETAMAL, Gustavo PUIATTI and Samuel BENVENISTES* Hospital Exequiel Gonzalez Cortes, Paed Urology, Santiago, CHILE - * Hospital Exequiel Gonzalez Cortes, Pathology Depart, Santiago, CHILE
PURPOSE Bladder augmentation (BA) using demucosalized sigmoid colon (DSC) is associated with a decrease in mucus production. The aim of this study is to determinate the histological changes at various times after augmentation.
MATERIAL AND METHODS 24 specimens were taken from 20 patients who had undergone BA using DSC over 7 years (1998-2005). Biopsy was done in 3 different points of the augmented patch (9-12-3 o’clock). In children with a closed bladder neck a trans-vesical trochar was used. Histopathology (HþE) results were compared at different post augment
periods. The biopsies were compared with 2 cases of BA where sigmoid colon without demucosalisation was used (SCWD)(>10 years).
with atrophic glands. In the group >36 months the patch was mostly covered with intestinal mucosa with atrophic glands surrounded by fibrotic tissue and no signs of urothelium. In the 2 cases of BA with SCWD the intestinal glands were normal.
RESULTS The mean age at biopsy was 12 years (range 4e15y) with a mean post BA of 39 months (range 12e84m). On endoscopy there were 2 groups; one (<3 years) where the intestine was covered by urothelium with some small areas of intestinal mucosa (‘‘islets’’) (n¼9), and the other >3 years where the patch was covered completely with intestinal mucosa (n ¼ 15). The histology of the early biopsies (<36 months) showed urothelium with some small ‘‘islets’’ of sigmoid mucosa, but
CONCLUSIONS This study has demonstrated regrowth of sigmoid colon mucosa, which may begin from intestinal mucosa remnants (‘‘islets’’), noted on specimens taken <3 years after BA. However the glandular structure is atrophic in those patches, and was normal in those cases augmented with non-demucosalized sigmoid. This may explain the reduced mucus production in those patients augmented with DSC.